TissueCultureMedia

Weusetwodifferentkindsofmedia.MostcellsaregrowninDMEM.AfewlymphoidcelllinesaregrowninRPMI.CellsgrowninDMEMmustbegrownina10%CO2atmosphere.Asaresult,mostofourincubatorsaresetforthis.Incontrast,cellsgrowninRPMImustbegrownina5%CO2atmosphere.Wehaveoneincubatorsetupforthis.IfyougrowcellsinRPMIin10%CO2,themediumwillbetooacidic(yellow).

DMEM.

ThisisanabbreviationforDulbecco/VogtmodifiedEagle"s(HarryEagle)minimalessentialmedium.Weusecommercially-madeDMEMasthemediumformostcells.DMEMdiffersfromtheoriginalMEMinthatitcontainsapproximately4timesasmuchofthevitaminsandaminoacidspresentintheoriginalformula,andsome(2to4fold)moreglucose.Italsocontainsironandafewotheroddments.ThesemodificationsweremadebyMargueritesoshedidnothavetocomeinonSundaytochangethemediumonhercells.Mostkindsofcells--human,monkey,hamster,rat,mouse,chicken--growwellinthismedium.

RPMI.

ThisisanabbreviationforRoswellParkMemorialInstitutemedium.HumanlymphoidcellsaretrADItionallygrowninRPMImedium.ThisismadefromamixbyMargie.Thismediumcontainsagreatdealofphosphateandisformulatedfor5%CO2.Mediumisalwayssupplementedwithserum.MostcelllinesaregrowninDMEMsupplementedwitheither10%calfserumor10%fetalcalfserum.Transformedcellsusuallygrowwellincalfserum.Normal9non-transformed)cellssometimesrequirefetalcalfserum,whichisricher.Someclonesof3T3cellsdonoticeablybetterinfetalcalfserum.Someveryfastidiouscellsaregrownin20%fetalcalfserum.Duetothecostandscarcityoffetalcalfserum,somecellsaregrowninhorseseruminstead.Themediumofsomelymphoidcellsissupplementedwith5x10-5M2-mercaptoethanol.Itdoesnotmatterwhetherthishasundergoneoxidation.Somecellsbenefitfromtheinclusionof2mMglutamine,2,5or10%tryptosephosphatebroth,2mMpyruvate,ornon-essentialaminoacidsintheirmedia.Forexample,Mary-AnnCampbellalwaysputspyruvateandnon-essentialaminoacids,inadditiontomercaptoethanol,inthemediumofherBcells.